| Autor: |
Matsuike D; Graduate School of Science, Osaka Metropolitan University, Osaka, Japan., Tahara YO; Graduate School of Science, Osaka Metropolitan University, Osaka, Japan.; OCU Advanced Research Institute for Natural Science and Technology, Osaka Metropolitan University, Osaka, Japan., Nonaka T; Graduate School of Science, Osaka City University, Osaka, Japan., Wu HN; Graduate School of Science, Osaka City University, Osaka, Japan., Hamaguchi T; Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Miyagi, Japan., Kudo H; Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Meguro, Tokyo, Japan.; Graduate School of Science, Technology and Innovation, Kobe University, Nada, Kobe, Japan., Hayashi Y; Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Meguro, Tokyo, Japan.; Environmental Science Center, The University of Tokyo, Bunkyo, Tokyo, Japan., Arai M; Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Meguro, Tokyo, Japan.; Department of Physics, Graduate School of Science, The University of Tokyo, Meguro, Tokyo, Japan., Miyata M; Graduate School of Science, Osaka Metropolitan University, Osaka, Japan.; OCU Advanced Research Institute for Natural Science and Technology, Osaka Metropolitan University, Osaka, Japan. |
| Abstrakt: |
Mycoplasma mobile is a fish pathogen that glides on solid surfaces by means of its own gliding machinery composed of internal and surface structures. In the present study, we focused on the function and structure of Gli123, a surface protein that is essential for the localization of other surface proteins. The amino acid sequence of Gli123, which is 1,128 amino acids long, contains lipoprotein-specific repeats. We isolated the native Gli123 protein from M. mobile cells and a recombinant protein, rGli123, from Escherichia coli. The isolated rGli123 complemented a nonbinding and nongliding mutant of M. mobile that lacked Gli123. Circular dichroism and rotary-shadowing electron microscopy (EM) showed that rGli123 has a structure that is not significantly different from that of the native protein. Rotary-shadowing EM suggested that Gli123 adopts two distinct globular and rod-like structures, depending on the ionic strength of the solution. Negative-staining EM coupled with single-particle analysis revealed that Gli123 forms a globular structure featuring a small protrusion with dimensions of approximately 15.7, 14.7, and 14.1 nm for the "height," major axis and minor axis, respectively. Small-angle X-ray scattering analyses indicated a rod-like structure composed of several tandem globular domains with total dimensions of approximately 34 nm in length and 6 nm in width. Both molecular structures were suggested to be dimers, based on the predicted molecular size and structure. Gli123 may have evolved by multiplication of repeating lipoprotein units and acquired a role for Gli521 and Gli349 assembly. IMPORTANCE Mycoplasmas are pathogenic bacteria that are widespread in animals. They are characterized by small cell and genome sizes but are equipped with unique abilities for infection, such as surface variation and gliding. Here, we focused on a surface-localizing protein named Gli123 that is essential for Mycoplasma mobile gliding. This study suggested that Gli123 undergoes drastic conformational changes between its rod-like and globular structures. These changes may be caused by a repetitive structure common in the surface proteins that is responsible for the modulation of the cell surface structure and related to the assembly process for the surface gliding machinery. An evolutionary process for surface proteins essential for this mycoplasma gliding was also suggested in the present study. |
